Improving the speed and efficiency of DNA sequencing methods is essential to the success of the Human Genome Program and to the success of many biotechnology companies such as CuraGen. DNA sequencing remains the rate determining step in much of genetic research. Our goal (and the subject of this proposal) is to dramatically increase the speed and efficiency of gel electrophoresis sequencing using fluorescently tagged chain terminators. We will investigate the upper throughput bounds of this method by (1) increasing optical format and efficiency, (2) massively increasing the lane density and hence the number of samples per run, and (3) improving data handling efficiency through software and dye coding strategies. Sensitivity and throughput increases of 20 fold each may be realized through implementation of optical improvements and a new separation technique. The optical spectroscopy, computation, mathematics, engineering and molecular biology backgrounds present at CuraGen combine with a strong consulting base to put us in a unique position to conceive and carry out this effort. If successful, the instrumental advances achieved in Phase I will be developed into a Phase II prototype capable of accepting four standard 96 well microtitration plates directly, with an immense commercial advantage over existing systems.